Exploring Modern Mechanism of Treating Diabetic Retinopathy and Coronary Heart Disease from Perspective of Blood Stasis Under Viewpoint of Treating Different Diseases with the Same Method / 中国实验方剂学杂志

Diabetic retinopathy（DR） and coronary heart disease（CHD） are both major chronic vascular complications that seriously jeopardize the health of the population and often occur together in clinical practice， it is of great clinical value to actively explore the association between the two in the process of disease development and methods of prevention and treatment of modern medicine and traditional Chinese medicine（TCM）. According to TCM， the heart and eyes physiologically communicate with each other by taking Qi， blood and veins as bridges， blood stasis obstructing collaterals is the common TCM etiology of DR and CHD， whose mechanism involves inflammation， oxidative stress and endothelial dysfunction. Promoting blood circulation and removing blood stasis plays an important role in the same treatment for different diseases and prevention and treatment of comorbidities， possibly by inhibiting the expression of interleukin-1β（IL-1β）， endothelin-1（ET-1） and hypoxia inducible factor-1α/vascular endothelial growth factor（HIF-1α/VEGF）， regulating phosphatidylinositol 3-kinases/protein kinase B/mammalian target of rapamycin（PI3K/Akt/mTOR） pathway， initiating adenosine monophosphate（AMP）-activated protein kinase/silent information regulator 1（AMPK/SIRT1） and nuclear transcription factor erythroid 2-related factor 2/heme oxygenase-1（Nrf2/HO-1） signaling pathways， inhibiting Hippo/Yes-associated protein（Hippo/YAP） signaling pathway， inhibiting mitochondrial permeability transition pore and anti-platelet agglutination for treating DR and CHD， which provides a multi-component， multi-pathway and multi-target selection strategies and ideas for the prevention and treatment of DR and CHD by TCM from a biological perspective. Based on this， subsequent studies should focus on constructing clinically relevant comorbidity models， conducting multicenter prospective studies， and fully utilizing artificial intelligence technology to gain a deeper understanding of the relationship between the two diseases， so as to elucidate the mechanism of promoting blood circulation and removing blood stasis in preventing and treating panvascular diseases.

Efficacy and safety evaluation of dabigatran etexilate in the treatment of elderly non-valvular atrial fibrillation / 药学实践杂志

Objective To explore the efficacy and safety of dabigatran etexilate in the treatment of elderly patients with non-valvular atrial fibrillation (NVAF), so as to provide reference for clinical treatment and rational drug use. Methods Eighty elderly patients with NVAF in The Seventh People's Hospital of Shanghai from December 2020 to June 2021, aged 65 to 80 years, were enrolled in a self-controlled study. Dabigatran etexilate 110 mg was given orally, twice a day (one in the morning and one in the evening) for 6 months. During the follow-up visit, the coagulation function indexes including APTT, TT, FIB and D-D were observed in patients taking medicine for 1 month and 6 months. Liver function indexes including ALT, AST and TBIL, renal function indicators including UREA and eGFR, and blood routine indexes including HGB and ESR, also were observed in patients taking medicine for 1 month and 6 months. Results Patients were compared at 1 month and 6 months after treatment with dabigatran and before: the difference of coagulation function indexes as APTT and TT was statistically significant (P<0.05);There was no significant difference in liver function index, renal function index and blood routine index (P>0.05); A total of 25 adverse events occurred, and the incidence rate of adverse reactions was 31.25%, there was no adverse events such as serious hemorrhage，life threatening and organ failure hemorrhage occurred．Conclusion Dabigatran has good efficacy and safety in the treatment of the elderly NVAF.

UHRF1/DNMT1-MZF1 axis-modulated intragenic site-specific CpGI methylation confers divergent expression and opposing functions of PRSS3 isoforms in lung cancer

As confusion mounts over RNA isoforms involved in phenotypic plasticity, aberrant CpG methylation-mediated disruption of alternative splicing is increasingly recognized as a driver of intratumor heterogeneity (ITH). Protease serine 3 (PRSS3), possessing four splice variants (PRSS3-SVs; PRSS3-V1-V4), is an indispensable trypsin that shows paradoxical effects on cancer development. Here, we found that PRSS3 transcripts and their isoforms were divergently expressed in lung cancer, exhibiting opposing functions and clinical outcomes, namely, oncogenic PRSS3-V1 and PRSS3-V2 versus tumor-suppressive PRSS3-V3, by targeting different downstream genes. We identified an intragenic CpG island (iCpGI) in PRSS3. Hypermethylation of iCpGI was mediated by UHRF1/DNMT1 complex interference with the binding of myeloid zinc finger 1 (MZF1) to regulate PRSS3 transcription. The garlic-derived compound diallyl trisulfide cooperated with 5-aza-2'-deoxycytidine to exert antitumor effects in lung adenocarcinoma cells through site-specific iCpGI demethylation specifically allowing MZF1 to upregulate PRSS3-V3 expression. Epigenetic silencing of PRSS3-V3 via iCpGI methylation (iCpGIm) in BALF and tumor tissues was associated with early clinical progression in patients with lung cancer but not in those with squamous cell carcinoma or inflammatory disease. Thus, UHRF1/DNMT1-MZF1 axis-modulated site-specific iCpGIm regulates divergent expression of PRSS3-SVs, conferring nongenetic functional ITH, with implications for early detection of lung cancer and targeted therapies.

Analysis of gene mutation type and frequency in children with thalassemia aged 0-18 years in Chengdu, Sichuan / 中华检验医学杂志

Objective:To analyze the type and frequency of thalassemia gene mutation in children aged 0 to 18 years in Chengdu.Methods:A total of 568 children from Chengdu, who were initially positive for thalassemia during screening from September 2018 to July 2021, were recruited. Among them, there were 308 males and 260 females. The type of mutation and distribution of α and β types of thalassemia in this cohort was analyzed utilizing PCR reverse dot blot.Results:Among the 568 children, 356 were genetically diagnosed as thalassemia, with a total positive rate of 62.68%. Among them, there were 140 cases of α-thalassemia with a positive rate of 24.65%, and 202 cases of β-thalassemia with a positive rate of 35.56%. There were 14 carriers of α-complex β-thalassemia gene, and the positive rate was 2.46%. Among these cases, the types of α-thalassemia gene mutation were mainly αα/--sea (79.29%), αα/-α3.7 (7.86%), and-α3.7/--sea (7.14%) genotypes, accounting for 94.29% of all types. In the 202 β-thalassemia patients, 199 heterozygous mutations were identified, mainly including cd17(A?T) (36.13%), cd41-42(-TCTT) (32.68%), IVS-2-654(C?T) (20.79%), and accounting for 88.61% of all types of gene mutation, and 3 compound heterozygous mutations were detected. α-complex β-thalassemia was detected in 14 patients, including cd41-42(-TCTT)/-α3.7, VS-2-654(C?T)/--sea, cd17(A?T)/-α3.7 and cd41-42(-TCTT)/--sea, which accounting for 57.14% of all types of gene mutation. Our results showed that there is no sex difference between α and β thalassemia in Chengdu area, whereas the prevalence of α combined with β thalassemia is higher in males ( P=0.003). Conclusions:The type of α-thalassemia mutation in Chengdu is mainly αα/--sea, whereas β-thalassemia with cd17 (A?T) mutations and α-complex β-thalassemia are more frequent in males. This study provides a reference for the formulation of prevention and control strategies for thalassemia in Chengdu.

Interpretation of pharmacokinetic-based criteria for supporting alternative dosing regimens of programmed cell death receptor-1 (PD-1) or programmed cell death-ligand 1 (PD-L1) blocking antibodies for treatment of patients with cancer guidance for industry / 中国临床药理学与治疗学

In recent years, modeling and simulation technology based on pharmacometrics has received increasing attention in the development of innovation drugs. In August of 2021, FDA issued a guidance named Pharmacokinetic-Based Criteria for Supporting Alternative Dosing Regimens of Programmed Cell Death Receptor-1 (PD-1) or Programmed Cell Death-Ligand 1 (PD-L1) Blocking Antibodies for Treatment of Patients with Cancer Guidance for Industry, claiming the necessity of using population PK-based simulation method for the optimization of dosing regimens, and the corresponding implementation standards. This article first summarized the existing therapeutic regimens of PD-1/PD-L1 blocking antibodies in clinic as well as the main content of the guidance, and then cited some actual examples where population PK-based simulation method did contribute to the approval of the alternative dosing regimens. Besides, some critical considerations for the dosing regimen optimization of PD-1/PD-L1 blocking antibodies were also analyzed. In our view, this guidance would have positive impacts on the development of PD-1/PD-L1 blocking antibodies in the future. We hope that this article may provide some references for the colleagues in China.

Development and performance test of the environment scale of unintentional injury in the home for children aged 0-6 years old in urban area of China / 中华预防医学杂志

Objective@#To develop the environment scale of unintentional injury in the home for children aged 0-6 years living in urban area of China, and test its validity and reliability.@*Methods@#The content of the environment scale was established through the literature review, expert consultation and pilot study. A total of 1 104 children aged 0-6 years in urban area of Changsha were enrolled in this study by using a multi-stage stratified cluster random sampling method. The questionnaire was used to collect the basic information of children, the incidence of unintentional injury and the status of home environment. The reliability of the scale was tested by using Cronbach′s α coefficient and split-half reliability coefficient. The content validity and construct validity were tested by using Pearson correlation analysis and factor analysis. All children were divided into two groups according to the incidence of unintentional injury in the home and the discrimination validity of the scale was tested by using t-test.@*Results@#The scale had 54 items in 6 dimensions. The number of eligible questionnaires was 1 074, including 554 (51.6%) from boys and 519 (48.3%) from children under 3 years old. The incidence rate of unintentional injury and in-home injury was 18.34% (197 children) and 10.71% (115 children). The internal consistency reliability (Cronbach′s α) coefficient for the scale was 0.87 and the split-efficacy reliability coefficient was 0.82, both meeting the standard of reliability above 0.70. The Pearson correlation coefficient between each dimension and the whole scale ranged from 0.53 to 0.84 (all P values <0.001). The common factor cumulative variance contribution rate of the scale was 58.34%. There were 54 items with factor loadings greater than 0.30. The root mean square error of approximation, comparative fit index and goodness-of-fit index were 0.07, 0.61 and 0.71, respectively. The score of scale in children with injury was significantly higher than that in children without injury (P=0.022).@*Conclusion@#The validity and reliability of the environment scale for unintentional injury in the home for children aged 0-6 years old in the urban area of China are good.

Advances in the study of Parkinson′s tremor / 中华神经科杂志

Parkinson′s disease is a neurodegenerative disease with early prominent death of dopaminergic neurons in the substantia nigra pars compacta. The resultant dopamine deficiency within the basal ganglia leads to a movement disorder characterized by classical parkinsonian motor symptoms. At present, the mechanism of Parkinson′s disease tremor has not been fully elucidated, and there are different hypotheses. Based on Duval′s hypothesis of "finger-switch-dimmer" on the pathogenesis of tremor in Parkinson′s disease, and combined with clinical practice, this review comprehensively expounds how tremor in Parkinson′s disease is generated under this model, and introduces the classification, diagnosis and treatment of tremor in Parkinson′s disease, so as to provide new ideas for the treatment of tremor in Parkinson′s disease in the future.

Doxorubicin-eluting beads vs conventional TACE for hepatocellular carcinoma: a Meta-analysis / 中国医学物理学杂志

Objective To conduct a Meta-analysis for assessing the therapeutic efficacy and safety of doxorubicin-eluting bead transcatheter arterial chemoembolization (DEB-TACE) vs conventional TACE (C-TACE) in the treatment of hepatoeellular carcinoma (HCC).Methods The relevant studies on DEB-TACE and C-TACE for HCC were searched from PubMed,Web of Science and CK1N databases.The tumor responds and adverse events in the selected studies were analyzed with RevMan5.0 statistical software.Moreover,publication bias was assessed by a funnel plot.Results Thirteen studies with 1 325 HCC patients were finally included in this analysis.The tumor response to DEB-TACE was better than that to C-TACE (OR:2.28;95％CI [1.75,2.96];P＜0.001 0).The risk of adverse events in DEB-TACE group was also lower than that in C-TACE group (RR:0.56;95％ CI [0.32,0.97];P=0.04).Conclusion Compared with C-TACE,DEB-TACE is more effective and has lower risk of adverse events in the treatment of HCC.

Effect of alveolar macrophages phagocytosis on prognosis in patients with acute respiratory distress syndrome caused by abdominal infection: a multicenter study / 中华危重病急救医学

Objective To investigate the effects of alveolar macrophage phagocytosis on prognosis in patients with acute respiratory distress syndrome (ARDS) caused by abdominal infection. Methods ARDS patients caused by severe intra-abdominal infection admitted to intensive care unit (ICU) of Tianjin Fourth Central Hospital, Tianjin Nankai Hospital, Tianjin First Central Hospital and Tianjin Fifth Central Hospital from June 2016 to March 2018 were enrolled. The gender, age, acute physiology and chronic health evaluationⅡ(APACHEⅡ) within 24 hours of admission, neutral red phagocytosis and alkaline phosphatase activity of macrophages in bronchoalveolar lavage fluid, the length of ICU stay, total hospitalization time, hospitalization expenses, and prognosis were recorded. According to the prognosis, the patients were divided into death group and survival group, and the parameters were compared between the two groups. Pearson test was used to analyze the correlation between neutral red phagocytosis function of macrophages and alkaline phosphatase activity and other indicators. The prognosis was analyzed by binary Logistic regression combined with neutral red phagocytosis and alkaline phosphatase activity in patients, and the predictive value of both subjects on prognosis was analyzed by the receiver operating characteristic (ROC) curve. Results Twenty patients were enrolled in the study, with 8 in the death group and 12 in the survival group. Compared with the survival group, the death group was older (years old: 58.50±14.86 vs. 46.67±13.40), APACHEⅡ score was higher (21.50±3.93 vs. 13.58±4.12), neutral red phagocytosis ability and alkaline phosphatase activity of alveolar macrophages were significantly decreased (A value:0.265±0.050 vs. 0.338±0.016; μmol/L: 12.06±1.24 vs. 17.96±3.90), and the length of ICU stay was significantly longer (days: 22.00±14.59 vs. 11.50±3.17), hospitalization cost was significantly increased (10 thousand Yuan:24.17±11.02 vs. 13.44±3.53), the total hospitalization time was shorter (days: 25.25±15.01 vs. 35.67±8.58), and the difference was statistically significant (all P < 0.05). There was no significant difference in gender between the survival group and the death group [male (case): 8 vs. 6, P > 0.05]. The neutral red phagocytosis ability of alveolar macrophages in ARDS patients caused by abdominal infection was negatively correlated with age, APACHEⅡ score and the length of ICU stay (r value was -0.328, -0.572, -0.809, respectively, all P < 0.05); alkaline phosphatase activity was negatively correlated with age, APACHEⅡ score, the length of ICU stay and hospitalization expenses (r value was -0.334, -0.583,-0.470, -0.517, respectively, all P < 0.05). Binary Logistic regression analysis showed that neutral red phagocytosis [odds ratio (OR) = 0.596, 95% confidence interval (95%CI) = 0.212-0.997] and alkaline phosphatase activity (OR = 0.573, 95%CI = 0.339-0.968) were the influencing factors of prognosis (both P < 0.05). ROC curve analysis showed that the AUC of neutral red phagocytosis ability for prognosis of ARDS patients caused by abdominal infection was 0.948, and the sensitivity and specificity were 91.7% and 87.5% when the off-cut value was 0.317. The AUC of alkaline phosphatase for the prognosis of ARDS patients caused by abdominal infection was 0.813; when the cut-off value was 19.72 μmol/L, the sensitivity was 75.0%, and the specificity was 87.5%. Conclusion The alveolar macrophage phagocytosis dysfunction in ARDS patients caused by severe abdominal infection was not only related to the severity of the disease, but also increased the medical burden of patients, and significantly affected the mortality of such patients.

Advances in the study of Parkinson′s tremor / 中华神经科杂志

Parkinson′s disease is a neurodegenerative disease with early prominent death of dopaminergic neurons in the substantia nigra pars compacta. The resultant dopamine deficiency within the basal ganglia leads to a movement disorder characterized by classical parkinsonian motor symptoms. At present, the mechanism of Parkinson′s disease tremor has not been fully elucidated, and there are different hypotheses. Based on Duval′s hypothesis of "finger?switch?dimmer" on the pathogenesis of tremor in Parkinson′s disease, and combined with clinical practice, this review comprehensively expounds how tremor in Parkinson′s disease is generated under this model, and introduces the classification, diagnosis and treatment of tremor in Parkinson′s disease, so as to provide new ideas for the treatment of tremor in Parkinson′s disease in the future.

Role of interictal epileptiform discharge in reducing recurrent epilepsy after withdrawal of antiepileptic drugs / 中华老年心脑血管病杂志

Objective To study the role of interictal epileptiform discharge (IED) in reducing recurrent epilepsy after withdrawal of antiepileptic drugs (AED).Methods One hundred epilepsy pa tients with no seizure for ≥2 years were divided into IED group (n=51) and IED free group (n=49) according to the classification of epilepsy seizure developed by the International Association for the Prevention of Epilepsy in 1981.The patients were further divided into elderly group (n=21) and non-elderly group (n=79) and were followed up for at least 1 year by return visit or telephone.Results No significant difference was found in the incidence of IED in the 100 epilepsy patients with different types of seizure,such as myospasm,simple partial seizure and ≥2 seizures.However,the incidence of IED was significantly higher than that of myotonia,myospasm,absence and simple partial seizure (P＜0.05).Epilepsy recurred in 37 patients (72.5％) of IED group and in 16 patients (32.7％) of IED-free group.Logistic regression analysis showed that the course of epilepsy and IED were the risk factors for recurrent epilepsy after withdrawal of AED (OR=1.165,95％CI:1.022-1.329,P=0.022;OR=2.794,95％CI:1.040-7.509,P=0.042) and the course of epilepsy was longer in elderly group than in non-elderly group (10.10±7.55 years vs 5.97±4.04 years,P=0.001).Conclusion The seizure type and course of epilepsy are the relia ble predictors of recurrent epilepsy in patients with no seizure for a long time after withdrawal of AED,and are thus of clinical significance.

Anti-fatigue activity of fermented grain-containing blueberry anthocyanins / 军事医学

Objective To investigate the anti-fatigue activity of fermented grain-containing blueberry anthocyanins ( LANHE,LH) in mice.Methods Experiments were conducted in two phases .In the first phase , forty mice were randomly divided into four groups:control group(distilled water,dw) and three LH administration groups (8.75,17.5 and 35.08 ml/kg body mass).In the second phase, mice were randomly divided into two groups:high-dose LH group (35.08 ml/kg body mass) and control group (dw 35.08 ml/kg body mass).After four weeks, a forced swimming test was performed and the biochemical parameters related to fatigue were examined .Results and Conclusion The administration groups showed a significant increase of swimming time to exhaustion compared with the control group , especially the high-dose group ( P0.05).LH could significantly increase the liver glycogen contents and decrease the serum contents (P<0.05).These data indicate that LH has anti-fatigue activity and can elevate the exercise tolerance in mice .

Clinical features and individualized treatment of 39 cases of adefovir dipivoxil-induced renal hypophosphatemia and osteomalacia / 中华传染病杂志

Objective To investigate the clinical features,early diagnosis and individualized treatment of renal hypophosphatemia and osteomalacia induced by adefovir dipivoxil (ADV) in patients with chronic hepatitis B (CHB).Methods Thirty-nine CHB or hepatitis B virus (HBV)-related cirrhosis patients of renal hypophosphatemia and osteomalacia induced by ADV were consecutively collected.The clinical features were analyzed and treatment outcome was followed up.Results The mean age of the 39 patients was 54 (27-71) years old.There were 26 male and 13 female patients,and 19 patients with cirrhosis.The mean ADV treatment duration was 69 (range 18-116) months,and 31 patients were treated for 36-96 months.The mean serum phosphate was 0.68 (0.42-0.79) mmol/L.Twenty-six cases developed renal hypophosphatemic osteomaolacia,of which 14 had bone pain and 19 had abnormally elevated alkaline phosphatase (ALP).Three patients had increased serum creatinine and 24 patients had decreased estimated glomerular filtration rate (eGFR).After individualized treatment,patients gained normal serum phosphate in mean of 2.0 (range 0.5-6.0) months,and had bone pain remission in the mean of 0.8 (range 0.2-1.0) month and bone pain disappeared in the mean of 1.5 (range 0.5-5.0) months.Function indices of liver and kidney were improved gradually,and the bone mineral density examination improved slowly.Conclusions CHB and HBV-related cirrhosis patients treated with longterm ADV could develop renal hypophosphatemia and hypophosphatemic osteomalacia,which is partially reversible.Monitoring serum phosphate,creatinine and cystatin C is necessary during long-term ADV therapy.After confirmed diagnosis,withdrawal or dosage reduction of ADV,and switch to telbivudine or entecavir should be considered.Meanwhile,serum phosphate and HBV DNA level should be monitored.

Biocompatibility of magnetic chitosan microspheres / 中国组织工程研究

BACKGROUND:Chitosan modification for magnetic iron oxide nanoparticles can improve the agglomeration and stability of magnetic iron oxide nanoparticles, and in addition, it can be used for tumor hyperthermia and gene therapy in the future. OBJECTIVE:To explore the biocompatibility of magnetic chitosan microspheres which have the potential application in tumor hyperthermia and gene therapy. METHODS:Magnetic iron oxide nanoparticles were prepared using the modified chemical precipitation. Phacoemulsification method was used to add chitosan into magnetic iron oxide nanoparticles in the preparation of magnetic chitosan microspheres. Then, the fol owing tests were performed. (1) Cytotoxicity test:L-929 cells were cultured in 1640 medium, polyacrylamide monomer solution, 100%, 75%, 50%, 25%of magnetic chitosan microsphere extracts. (2) Hemolysis test:Magnetic chitosan microspheres leaching solution, saline and distil ed water were added. (3) Micronucleus test:Kunming mice were intraperitoneal y injected with magnetic chitosan microspheres suspension containing 5, 3.75, 2.5, 1.25 g/kg iron oxide magnetic fluid, cyclophosphamide and saline, respectively. RESULTS AND CONCLUSION:Magnetic chitosan microspheres of 200-300 nm in diameter had an increased dispersion effect. The result of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that the toxicity of the material on L-929 celllines belonged to no cytotoxicity. In the hemolysis test, the hemolysis rate of magnetic chitosan microspheres was 0.69%far less than 5%. In the micronucleus test, magnetic chitosan microspheres suspension did not cause DNA fragmentation and aneuploidy, and did not result in genetic toxicological effects produced by micronuclei. The magnetic chitosan microspheres did not appear with cacogenesis and mutagenesis. From the results, magnetic chitosan microspheres are a kind of high biocompatibility material.

Construction and identification of mammary expressional vector for cDNA of human lactoferrin / 生物工程学报

The aim of this study was to construct a mammary gland-specific expressional vector pBC1-hLF-Neo for Human Lactoferrin (hLF) gene and then investigate its expression in the mammary gland epithelium cells. The constructed vector contained the 6.2 kb long 5' flank regulation region including promoter, other elements and the 7.1 kb long 3' flank regulation region including transcriptional ending signal of a goat's beta-casein gene. A cassette of Neo gene was also inserted into the vector which gave a total length of 26.736 kb identified by restriction fragment analysis and partial DNA sequencing. The results revealed that the structure of the final constructed vector accords with the designed plasmid map. In order to analyze the bioactivity of the vector, we transfected the lined vector DNA into the dairy goat's mammary gland epithelium cells and C127 cells of a mouse's mammary epithelium by Lipofectamine. After selection with G418 for 8-10 days, G418-risistant clones were obtained. PCR analysis demonstrated that hLF gene cassette had been integrated into the genomic DNA of G418-risistant clones. After proliferation culture, the two kinds of transgenic cells were cultured in serum-free DMEM-F12 medium with prolactin, insulin and hydrocortisone- a medium capable of inducing recombinant hLF expression. RT-PCR, Western blotting and anti-bacteria bioactivity experiments demonstrated that the constructed mammary gland specific vector pBC1-hLF-Neo possessed the desirable bioactivity to efficiently express and could secrete hLF in both mammary gland cells and have the effect of E. coli proliferation inhibition. Paramount to everything, this study laid a firm foundation for preparing the hLF gene transgenic goat fetal-derived fibroblast cells.

Cryopreservation of mouse spermatogonial stem cells / 解剖学报

Objective To explore the conditions and methods for cryopreservation and proliferation of mouse spermatogonial stem cells (SSCs). Methods SSCs were isolated from six-day-old Kunming mouse using two-step enzymatic digestion and Percoll discontinuous density gradient centrifugation. Cells were frozen with different freezing medium and cooling rate. After thaw, they were cultured in mimimum essential medium alpha (MEMα) supplemented with 10% fetal calf serum (FCS) and 100μg/L glial cell line-derived neurotrophic factor (GDNF). The survived and proliferating SSCs were examined by WST-8 colorimetric assay. Alkaline phosphatase andreverse transcription-polymerase chain reaction (RT-PCR) were performed to confirm if the cultured 96 hours germ cells were still stem cells. Results The best method to cryopreserve SSCs is using cryoprotector containing 10% dimethyl sulfoxide(DMSO), 10% FCS, 0.07mol/L sucrose and 1℃/min cooling rate, and the viability of cells in this method is more than 84%;Although the cell viability in non-programmed freezing method is less than that in the programmed freezing method, it is a simple and effective cropreservation method for mouse SSCs. What is more, the anchoring time of SSCs in this method is 8-12 hours after thaw, SSCs begin to proliferate 24 hours later, and rapid proliferation appears on the 48 hours, colonies are composed by 20-25 cells in 96 hours, when SSCs proliferated nearly 5 times.Conclusion The culture condition we used is suitable for proliferation of frozen-thawed SSCs.

Cloning of human lysosomal acid beta-glucosidase gene and its expression in COS7 cells / 生物工程学报

In this study, we amplified human lysosomal acid beta-glucosidase (GlcCerase) gene by RT-PCR from human placenta, and analyzed the sequence of the PCR product cloned in pMD-19T. The gene identity was 99% comparable to that of the reported human GlcCerase cDNA sequence in GenBank. The GlcCerase gene digested with Xho I was subcloned into eukaryotic express vector pEGFP-C1 to generate recombinant expression vector pEGFP-GlcCerase. After identified the recombinant plasmid by restriction enzyme digestion, we transfected pEGFP-GlcCerase into COS7 cells by liposome. GlcCerase mRNA was expressed and the activity of GlcCerase was also detected in COS7 cells. This study would lay a foundation for the function of GlcCerase and its production by transgenic bioreactor.

Therapeutic effects of nano-magnetoliposomes containing As_2O_3 combined with magnetic fluid hyperthermia on xenograft cervical carcinoma / 中华物理医学与康复杂志

Objective To explore the inhibition effects and mechanics of nano-magnetoliposomes containing As_2O_3 (NMLA) combined with magnetic fluid hyperthermia on xenograft cervical carcinoma in nude mice. Methods After establishment of xenograft cervical carcinoma models in nude mice, nano-liposomes containing As_2O (NLA group), nano-magnetoliposomes (NML group) and NMLA (NMLA group) were injected into the xnograft cervical carcinoma, respectively, and all nude mice were exposed to AMF for three times. The inhibitive ratios (IR) of the tumors, apoptosis index (AI), proliferation index (PI) and the expression of Bcl-2 and Bax proteins were measured, as well as the function of liver and kidney. Results IRs in NLA, NML and NMLA groups were 57.06% (P0.05). The obvious necrosis was observed in the tumor tissues in NLA, NML and NMLA groups, but not in the peripheral tissues of the tumors and the visceras. Conclusion NMLA, without toxicity of liver and kidney, might be a desirable composite carrier of targeted treatment with chemotherapy and thermotherapy simultaneously.